(1) Field of the Invention
This invention relates to an apparatus for preparing large quantities of vaccine for mammals, and to the process for employing the substantially evacuated red blood cells of said mammals, thus providing a means of utilizing the erythrocytes in the vaccination of these animals.
(2) Description of the Prior Art
G. M. Ihler, et al. in Proceedings of the National Academy of Science of the U.S.A., Volume 70, pp. 2663-2666 (1973), used a large hypotonic dilution to encapsulate enzymes with a maximum encapsulation less than 1%. Later a one-third to one-fifth hypotonic dilution was used to encapsulate the enzyme uricase. In 1976 and 1977 other variations on the hypotonic dilution method have been used for other animal erythrocytes (see E. Ang, et al., Experimental Cell Research, Volume 104, pp. 430-434). Another method of encapsulation of substances in erythrocytes employs electrochemical shock [see F. Riemann, et al., Biochimica et Biophysica Acta, Volume 394, pp. 449-462 (1975)]; however, this method requires sophisticated equipment to generate exponentially decaying electric fields.
J. R. DeLoach et al. reported in 1977 a dialysis method for encapsulating enzymes and lipids in erythrocytes, where up to 45% of the added substances could be encapsulated. In that procedure, human erythrocytes at 70% to 80% hematocryt were dialyzed against 1 to 2 liters of distilled water for 2 hours. At 5 to 10 minute intervals the dialysis bag was removed and its contents mixed. Sufficient homogenous mixing was not achieved and it was difficult to prepare large quantities of resealed erythrocytes.
The prior art also discloses the use of dialysis to encapsulate enzymes with a maximum of about 30% encapsulated (see G. L. Dale, et al., Biochemical Medicine, Volume 18, pp. 220-225, 1977). More recently, M. Yaiuzimi, et al., in Cell, Volume 15, p. 245 (1978) employed a dialysis procedure for encapsulation of Immunoglobulin G (IgG) in erythrocyte ghosts. Although these authors reported the IgG concentrations inside the erythrocyte ghosts, no report on percent uptake was made.
The reports of the prior art indicate that there is yet much to be done to advance the state of the art. The present invention provides a step in that direction.